Anti-MTAP antibody [EPR22570-76]

11 product images

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  Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265)

  Lanes 1-4: Merged signal (red and green). Green - ab254265 observed at 32 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
  

  ab254265 Anti-MTAP antibody [EPR22570-76] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human MTAP knockout HeLa cell line ab265272 (knockout cell lysate Human MTAP knockout HeLa cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. ab254265 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265) at 1/1000 dilution

  Lane 1: Wild-type HeLa cell lysate at 20 µg

  Lane 2: MTAP knockout HeLa cell lysate at 20 µg

  Lane 3: HT-29 cell lysate at 20 µg

  Lane 4: A549 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 31 kDa

  Observed band size: 32 kDa

• 

  Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265)

  Lanes 1-4: Merged signal (red and green). Green - ab254265 observed at 32 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
  

  ab254265 Anti-MTAP antibody [EPR22570-76] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human MTAP knockout HeLa cell line ab265272 (knockout cell lysate Human MTAP knockout HeLa cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. ab254265 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265) at 1/1000 dilution

  Lane 1: Wild-type HeLa cell lysate at 20 µg

  Lane 2: MTAP knockout HeLa cell lysate at 20 µg

  Lane 3: HT-29 cell lysate at 20 µg

  Lane 4: A549 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 31 kDa

  Observed band size: 32 kDa

• 

  Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265)

  Blocking and dilution buffer: 5% NFDM/TBST.
  

  Exposure times:
  Lanes 1-4: 15 seconds; Lane 5: 10 seconds; Lane 6: 3.25 seconds.

  The expression molecular weight observed is consistent with what has been described in the literature (PMID:15492751).

  All lanes: Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265) at 1/1000 dilution

  Lane 1: Mouse skin lysate at 20 µg

  Lane 2: Mouse liver lysate at 20 µg

  Lane 3: Rat skin lysate at 20 µg

  Lane 4: Rat liver lysate at 20 µg

  Lane 5: RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg

  Lane 6: NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 31 kDa

  Observed band size: 32 kDa

• 

  Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265)

  Blocking and dilution buffer: 5% NFDM/TBST.
  

  Negative control: A549 (PMID: 17548352).

  The expression molecular weight observed is consistent with what has been described in the literature (PMID:15492751).

  Exposure times.

  Lanes 1-2: 10 seconds; Lane3: 5.5 seconds; Lanes 4-8: 3.25 seconds.

  All lanes: Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265) at 1/1000 dilution

  Lane 1: Human liver lysate at 20 µg

  Lane 2: Human placenta lysate at 20 µg

  Lane 3: HT-29 (human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

  Lane 4: HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

  Lane 5: HeLa (human epithelial cell line from cervix adenocarcinoma0 whole cell lysate at 20 µg

  Lane 6: Raji 9human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

  Lane 7: Caco-2 (human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

  Lane 8: A549 9human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

  Secondary

  Lanes 1 - 2: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution

  Lanes 3 - 8: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 31 kDa

  Observed band size: 32 kDa

• 

  Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265)

  anes 1- 4: Merged signal (red and green). Green - Anti-MTAP antibody [EPR6893] ab126770 observed at 32 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
  

  Anti-MTAP antibody [EPR6893] ab126770 Anti-MTAP antibody [EPR6893] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human MTAP knockout HeLa cell line ab265272 (knockout cell lysate Human MTAP knockout HeLa cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. Anti-MTAP antibody [EPR6893] ab126770 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265) at 1/1000 dilution

  Lane 1: Wild-type HeLa cell lysate at 20 µg

  Lane 2: MTAP knockout HeLa cell lysate at 20 µg

  Lane 3: HT-29 cell lysate at 20 µg

  Lane 4: A549 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 31 kDa

  Observed band size: 32 kDa

• 

  Flow Cytometry (Intracellular) - Anti-MTAP antibody [EPR22570-76] (ab254265)

  Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized A549 (human lung carcinoma cell line) (Left) anf H-29 (human colorectal adenocarcinoma cell line) (Right) cells labeling MTAP with ab254265 at 1/400 dilution (red) compared with Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).
  

  Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), at 1/2000 dilution was used as the secondary antibody.

  Negative control: A549 (PMID: 8971171.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR22570-76] (ab254265)

  Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling MTAP with ab254265 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in rat liver (PMID:15492751) is observed. Counter stained with hematoxylin.
  

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

  Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR22570-76] (ab254265)

  Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling MTAP with ab254265 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in mouse liver (PMID:15492751) is observed. Counter stained with hematoxylin.
  

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

  Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR22570-76] (ab254265)

  Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling MTAP with ab254265 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in stroma cells and no staining in tumor cells of human bladder cancer (PMID:27270441) is observed. Counter stained with hematoxylin.
  

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

  Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR22570-76] (ab254265)

  Immunohistochemical analysis of paraffin-embedded human breast tissue labeling MTAP with ab254265 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in human breast (PMID:26751376, 18712977) is observed. Counter stained with hematoxylin.
  

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

  Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-MTAP antibody [EPR22570-76] (ab254265)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) and HT-29 (human colorectal adenocarcinoma cell line) cells labeling MTAP with ab254265 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and nuclear staining in HT-29 cell line. The nuclear counter stain is DAPI (blue) Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 dilution (red).
  

  SEcondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

  Negative control: A549（PMID: 8971171.